Expression and significance of estrogen receptor α and β in prostate cancer and peri-cancer tissue

Objective To investigate the expression of estrogen receptor (ER) α and β in human prostate cancer (PC), peri-cancer tissue and benign prostatic hyperplasia (BPH) tissue, and to discuss the role of estrogen receptor in prostate cancer. Methods The expression of ERα and ERβ in PC (n=28), peri-cancer tissue (n=28) and BPH (n=29) were detected by immunohistochemistry with En vision method. The ERα and ERβ expression were compared among different tissues by chisquare. The relationship between ER expression and related clinicopathologic features was statistically analyzed by spearman rank collection. Results ERα was localized dominantly in the stromal cell of PC. There were significant differences of the expression of ERα in PC, peri-cancer tissue and BPH tissue (epithelial cell 0%, 14%, 24%, P＜0. 05; stromal cell 57%, 68%, 31%,P＜0. 05). ERβ was localized in both epithelial and stromal cell of PC. There were significant differences of the expression of ERβ in PC, peri-cancer tissue and BPH tissue (epithelial cell 39%, 64%, 29%, P＜0.01; stromal cell 50%, 75%, 79%, P＜0.05). There was a significant difference of the expression of ERβ in different Gleason scores of PC tissue. Conclusions ERα is localized in the stromal cell of PC tissue.ERβ is localized in both epithelial and stromal cell of PC tissue. The ERβ might be related to the tumor differentiation of PC.     

Expression and significance of estrogen receptor α and β in prostate cancer and peri-cancer tissue

Objective To investigate the expression of estrogen receptor (ER) α and β in human prostate cancer (PC), peri-cancer tissue and benign prostatic hyperplasia (BPH) tissue, and to discuss the role of estrogen receptor in prostate cancer. Methods The expression of ERα and ERβ in PC (n=28), peri-cancer tissue (n=28) and BPH (n=29) were detected by immunohistochemistry with En vision method. The ERα and ERβ expression were compared among different tissues by chisquare. The relationship between ER expression and related clinicopathologic features was statistically analyzed by spearman rank collection. Results ERα was localized dominantly in the stromal cell of PC. There were significant differences of the expression of ERα in PC, peri-cancer tissue and BPH tissue (epithelial cell 0%, 14%, 24%, P＜0. 05; stromal cell 57%, 68%, 31%,P＜0. 05). ERβ was localized in both epithelial and stromal cell of PC. There were significant differences of the expression of ERβ in PC, peri-cancer tissue and BPH tissue (epithelial cell 39%, 64%, 29%, P＜0.01; stromal cell 50%, 75%, 79%, P＜0.05). There was a significant difference of the expression of ERβ in different Gleason scores of PC tissue. Conclusions ERα is localized in the stromal cell of PC tissue.ERβ is localized in both epithelial and stromal cell of PC tissue. The ERβ might be related to the tumor differentiation of PC.     

瘦素对人乳腺癌裸鼠移植瘤雌激素受体α、βmRNA的调控作用

目的通过荷人乳腺癌裸鼠模型来进行在体研究外源性人类瘦素对移植瘤组织内雌激素受体(ER)a、β mRNA的调控作用.方法应用人乳腺癌MCF-7细胞制备荷人乳腺癌裸鼠模型,随机分成瘦素实验组(n=30)及0.9%的氯化钠溶液对照组(n=30).实验组采用瘤周皮下注射人重组瘦素连续15 d,对照组给予皮下注射相同剂量0.9%的氯化钠溶液.实时荧光PCR法扩增移植瘤组织内ERα、ERβ mRNA并进行相对定量分析.结果成功建立荷人乳腺癌裸鼠模型,并进行了瘤周皮下注射人类瘦素干预.ERα mRNA表达量在瘦素组高于0.9%的氯化钠溶液组(P＜0.01),ERβmRNA表达量在瘦素组低于0.9%的氯化钠溶液组(P＜0.01).结论通过瘤周皮下注射人类瘦素来干预荷人乳腺癌裸鼠是安全的.ERα与ERβ都可作为人类瘦素的作用靶点.外源性人类瘦素可以上调人乳腺癌裸鼠异种移植瘤内ERαmRNA的表达,下调ERβ mRNA的表达.

Abstract：

Objective To investigate the different effect of exogenous leptin on estrogen receptor α,β mRNA in human breast tumor tissue in nude mice xenograft models. Methods We made nude mice xenograft models of MCF-7 human breast cancer cells cultured in vitro, then divided them into experimental group of]eptin( n = 30)and control group of normal saline( n = 30)randomly. The models of experimental group were injected subcutaneously the recombinant human leptin for 15 consecutive days, the models of control group were injected subcutaneously the same dose of normal saline. A real- time quantitative RT- PCR assay was developed to quantify the expression of estrogen receptor α, β mRNA in tumor tissue, using the relative quantitative analysis. Results The leptin-intervened nude mice xenograft models were safely established. The relative quantitation of estrogen receptor α mRNA was significantly higher in the leptin group than in the normal saline group ( P ＜ 0.01 ), the relative quantitation of estrogen receptor β mRNA was significantly lower in the leptin group than in the normal saline group ( P ＜ 0. 01 ). Conclusion The nude mice xenograft models can be safely intervened with human leptin by subcutaneous injection around tumor.Estrogen receptor is one of the targets of leptin in the progress of breast cancer. Exogenous human leptin can up- regulate the expression of estrogen receptor α and down- regulate the expression of the estrogen receptor βin nude mice xenograft models of human breast tumor.     

雌激素受体-β在膀胱尿路上皮癌中的表达及其临床意义

目的 观察雌激素受体β(ER-β)在膀胱癌中的表达及其与临床病理特征的关系,探讨其临床意义.方法 采用免疫组织化学SP法检测ER-β蛋白在146例膀胱尿路上皮癌和25例正常膀胱黏膜中的表达.结果 ER-β蛋白在膀胱癌组织中阳性表达率为44.5%,在正常膀胱黏膜组织阳性表达率为20.0%,差异有统计学意义(P＜0.05).ER-β的表达率随着膀胱癌病理分级和临床分期的升高而增高(P＜0.05),而且与患者性别明显相关,女性阳性率显著高于男性患者(P＜0.05).结论 ER-β在膀胱癌组织中表达有明显的性别差异,且与膀胱癌的分化和浸润性进展密切相关,提示ER-β蛋白可能在膀胱癌的发生发展中起重要作用.

Abstract：

Objective To investigate the expression of estrogen receptor beta (ER-β) in bladder urothelial carcinoma and to explore its clinical significance.Methods Immunohistochemical SP method was employed to detected the expression of ER-β in bladder cancer tissue ( 146 cases) and the normal controls (25 cases).In addition,clinicopathological data of them were analyzed.Results The positive expression rate of ER-β in bladder cancer tissue was 44.5%,but low positive expression rate (20.0% )was found in normal bladder tissue.The expression of ER-β was markedly associated with tumor pathological grade and clinical stage ( P＜0.05).Moreover,the expression of ER-β was observed more frequently in female (59.6%) than male (37.4%) (P＜0.05 ).Conclusion The sex difference in expression of ER-β is associated with differentiation and invasion in bladder urothelial carcinoma,it implies that ER-β might play important roles in the development and progression of bladder cancer.     

Quantified gene expression levels for phase Ⅰ/Ⅱ metabolizing enzyme and estrogen receptor levels in benign prostate from cohorts designated as high-risk （UK） versus low-risk （India）for adenocarcinoma at this organ site：a preliminary study

Risk of clinically significant prostate adenocarcinoma （CAP） varies worldwide,although there is a uniform prevalence of latent disease. A hormone-responsive tissue,the prostate possesses the metabolizing capacity to biotransform a variety of environmental procarcinogens or endogenous hormones. Whether such metabolizing capacity or estrogen receptor （ER） status underlies these demographic differences in susceptibility to CaP remains unclear. With appropriate ethical permission,verified-benign tissues were obtained following transurethral resection of the prostate from a high-risk region （n = 12 UK-resident Caucasians） and a typically low-risk region （n = 14 India-resident Asians）. Quantitative gene expression analysis was employed for cytochrome P450 （CYP）1B1,N-acetyltransferase （NAT）1,NAT2,catechol-O-methyl transferase （ COMT）,sulfotransferase （ SULT） 1A1,ERα,ERβ and aromatase （CYP To quantify the presence or absence of CYP1B1,ERα or ERβ,and to identify ther in situ localization,immunohistochemistry was carried out. The two cohorts had reasonably well-matched serum levels of prostate-specific antigen or hormones. Expression levels for the candidate genes investigated were similar.However,clear differences in protein levels for CYP1B1 and ERβ were noted. Staining for CYP1B1 tended to be nuclear-associated in the basal glandular epithelial cells,and in UK-resident Caucasian tissues was present at a higher （P = 0.006） level compared with that from India-resident Asians. In contrast,a higher level of positive ERβ staining was noted in prostates from India-resident Asians. These study findings point to differences in metabolizing capacity and ER status in benign prostate tissues that might modulate susceptibility to the emergence of clinically significant CaP in demographically distinct populations.     

雌二醇通过雌激素受体α介导调控Cyclin D1在小鼠前列腺平滑肌细胞中的表达

目的 观察雌激素受体α(ERα)在17β-雌二醇(E2)促进小鼠前列腺平滑肌细胞(PSMC)增殖中的作用,探讨E2对PSMC的作用机制.方法 构建pGSadeno-ERα腺病毒载体,体外转染原代培养的小鼠前列腺平滑肌细胞(PSMC),将实验细胞分组:实验组;ERα基因下调组,细胞转染ERα干扰序列腺病毒;阴性对照组,细胞转染无义序列腺病毒;正常对照组,正常的小鼠PSMC.各组分别加入1×10-8mol/L E2,72 h后流式细胞仪检测细胞增殖,Western blot检测细胞内Cyclin D1基因表达.结果 E2作用细胞72 h后,实验组细胞G0/G1期的比率(71.76±1.78)%显著高于阴性对照组(51.73±1.86)%和正常对照组(47.90±0.79)%,差异有统计学意义(P＜0.05);相对应的PI值:实验组(28.24±1.78)显著低于阴性对照组(47.27±1.86)和正常对照组(52.10±0.79)比较,差异有统计学意义(P＜0.05);实验组细胞内Cyclin D1基因表达水平显著低于对照组.结论 雌激素在对前列腺平滑肌细胞的生物学效应中,通过雌激素受体α的介导调控了细胞周期素Cyclin D1的表达,从而促进细胞的增殖.

Abstract：

Objective To investigate the role of estrogen receptor alpha (Erα) in estradiol-induced proliferation of mouse prostatic smooth muscle cells (PSMCs) and explore a new target for benign prostazic hyperplasia (BPH). Methods Adenoviral vectors with Erα-shRNA or Ctrl-shRNA were transfected into mouse PSMCs. The cells were divided into 3 groups: experimental group ( cells transfected with pGSadeno-Erα), negative control group (cells transfected with pGSadeno-Ctrl) and normal control group (normal mouse PSMCs). The expression of Cyclin DI was detecrted by Western blotting. Cell cycle was analyzed by using flow cytometry. Results After treatment in the presence of 1 10-8 mol/L 17β-estradiol (E2) for 72 h, the percentage of cells in the G0/G1 phase was (71.76 1.78)％ in experimental group, which was significantly higher than in negative control group (51.73 ± 1.86)％ and normal control group (47.90±0.79)％, and PI in experimental group (28.24±1.78) was prominently lower than in negative control group (47.27±1.86) and normal control group (52.10±0.79). The expression level of Cyclin D1 in experimental group was conspicuously lower than in control groups. Conclusion E2 regulates the expression of Cyclin D1 via Erα in PSMCs, which promotes cell proliferation.     

XAGE-1 b基因在良性前列腺增生症与前列腺癌中的表达及其意义

目的 观察XAGE-1b基因在良性前列腺增生症和前列腺癌中的表达,探讨其数值对良性前列腺增生症和前列腺癌各项指标的临床意义.方法 运用实时荧光定量聚合酶链反应(PCR)方法检测38例前列腺癌病理组织以及40例良性前列腺增生症组织XAGE-1b的基因表达水平.结果 XAGE-1b在前列腺癌组织中表达值为8.299 50±0.97116,在良性前列腺增生症组织中表达值为3.007 80±0.91600,差异有统计学意义(P＜0.05).XAGE-1b表达值随着Gleason评分、临床分期和肿瘤恶性程度升高而表达增强(P＜0.05).结论 XAGE-1b基因高表达值有助于前列腺癌的诊断和指导前列腺癌的恶性程度分期.

Abstract：

Objective To study the XAGE-1 b mRNA expression in prostate cancer (PCa) tissues and benign prostate hyperplasia (BPH) tissues,and explore the diagnostic values of XAGE-1 b mRNA expression level in PCa and BPH.Methods A sensitive,real-time quantitative polymerase chain reaction (PCR) assay was developed to compare the expression difference of XAGE-1b mRNA in PCa and BPH tissues,by testing 38 samples of PCa and 40 samples of BPH.Results The expression level of XAGE-1 b in PCa tissue was 8.299 50 ± 0.971 16,and 3.007 80 ± 0.916 00 in BPH tissue ( P＜0.05 ).The XAGE-1 b expression levels were increased with the increase of the Gleason score,clinical stage and malignant grade (P＜0.05).Conclusion The high expression of of XAGE-1 b mRNA can afford a reliable and helpful information for diagnosis of PCa and BPH,and PCa malignant grade.     

Antagonism of estrogen-mediated cell proliferation by raloxifene in prevention of ageing-related prostatic hyperplasia

Estrogen has important roles in the initiation and development of benign prostatic hyperplasia （BPH）. Regulators of the estrogen receptor （ER） are tissue- and cell-specific. We evaluated the effect of estrogen antagonist, raloxifene （Ral）, on the prevention and treatment of BPH by investigating its effect on the proliferation of two different prostate cell lines： a stromal cell line, WPMY-1, and a benign prostatic hyperplasia epithelial cell line, BPH-1. We additionally evaluated its effect on prostatic hyperplasia induced by estrogen and androgen in a rat model. The effect of Ral on the prevention of prostatic hyperplasia was analyzed by haematoxylin and eosin staining and quantitative immunohistochemistry （IHC） for proliferating cell nuclear antigen and α-smooth muscle actin. In vitro and in vivo, tamoxifen （Tam）, another anti-estrogen drug, and finasteride （Fin）, a drug for the clinical treatment of BPH, served as efficacy controls. The in vitro data showed that neither Ral nor Tam alone affected the proliferation of WPMY-1 and BPH-1, but both antagonized the effect of oestradiol in promoting the proliferation of the two cells. Results from the IHC staining of the rat prostates indicated that, similar to Tam and Fin, Ral inhibited the proliferation of stromal cells in vivo. Interestingly, in contrast to Tam, both Ral and Fin inhibited the proliferation of epithelial cells. Furthemore, Ral treatment much strongly decreased the number of prostatic acini and the surrounding layers of smooth muscle cells than Fin （P 〈 0.05）. Our data showed for the first time that Ral may have a role in the response of the rat prostate to selective ER modulators.     

乳腺癌组织中PSAmRNA的表达及其与ER亚型表达的关系

目的 探讨前列腺特异抗原（PSA）mRNA和雌激素受体（ER）β亚型在乳腺癌组织中的表达及其与不同雌激素受体亚型表达之间的关系.方法 检测35例乳腺癌和12例癌旁乳腺组织及10例乳腺纤维腺瘤组织中的PSA mRNA和ERβ mRNA表达;35例乳腺癌组织ERα和PR蛋白的表达,并分析乳腺癌组织PSA mRNA表达与雌激素受体亚型和PR表达之间的关系.结果 PSA mRNA在乳腺癌组织中的表达水平明显低于癌旁乳腺组织和乳腺纤维瘤组织（P均=0.00）.ERβ mRNA在乳腺癌中的表达水平明显低于癌旁乳腺组织和乳腺纤维腺瘤组织（P均=0.00）.PSA mRNA表达阳性者ERβ mRNA表达水平明显低于PSA mRNA表达阴性者,差异有显著性（P=0.038）.ERα和PR阳性表达的乳腺癌组织中PSA mRNA表达高于ER和PR阴性者,差异有显著性（P=0.001,0.004）.结论 PSA和ERβ基因在乳腺癌中的表达下调.PSA可能是反映乳腺癌组织中功能性甾体类激素受体的一个重要指标.     

125碘标记17α-乙烯雌二醇-3-醋酸酯在 荷人乳腺癌裸鼠体内生物学分布△

Biodistribution of　¹²⁵I-labeled 17α-vinyestradiol-3-acetate (¹²⁵I-VE₂A)in nude mice bearing human breast cancer containing different estrogen receptor (ER) content was studied to understand the relation between this compound and ER and, consequently, to develop the ER imaging. Each mouse was injected with 92.5 kBq tracer from tail vein and then killed after two hours. The radioactivity uptake rate in one gram of tumor tissue and tissues from other vital organs were measured, and the radioactivity uptake ratio of tumor to non-tumor tissue was also measured. Results The radioactivity uptake rate and the radioactivity uptake ratio of tumor to non-tumor tissue in ER positive tumor (MCF-7) were much higher than those in ER negative tumor (MDA-MB-231). Conclusions This compound, IVE₂A has affinity to ER positive target organ or tumor and promise the probability to define the content and site of ER in vivo or in tumor.     

Androgen and prostatic stroma

Aim: To investigate the effect of androgen on the proliferation, differentiation and regression of canine prostatic stromal cells in vivo and human stromal cells in vitro. Methods: Twenty-two dogs, including 15 normal prostate dogs and 7 prostatic hyperplasia dogs, had their serum concentration of testosterone and estrodiol determined by radioimmunoassay before and after castration. The expression of androgen receptor (AR) and estrogen receptor (ER) in the prostate were analysed by immunohistochemistry and semi-quantitative RT-PCR before and after castration. Light microscopy, transmission electron microscopy and TUNEL assay were carried out successively before and after castration to evaluate the prostatic histomorphology. In vitro serum-free cell cultures from human prostatic stroma were established and exposed to dihydrotestosterone (DHT). The proliferation of the cell culture was detected by MTT assay. The expression of TGFβ, bFGF, AR, and smooth muscle cell (SMC) specific proteins (myosin and/or sm     

胰腺癌中雌激素受体α与β的表达

目的：检测胰腺癌中雌激素受体α、β（ERα、ERβ）蛋白和mRNA的表达。方法：采用免疫组化和RT-PCR检测50例胰腺癌和20例乳腺癌患者ERα、ERβ蛋白的表达以及ERα、ERβ mRNA的表达情况。结果：胰腺癌ERα蛋白的阳性率为32%,ERβ蛋白的阳性率为38%。胰腺癌ERα mRNA/ERβ mRNA的比值明显低于ER（ ）和ER（-）的乳腺癌。结论：ERβ在胰腺癌的发生、发展中可能起重要的作用。     

HIF-1α、P-gp在胰腺癌中的表达及其临床病理学的意义

目的 检测HIF-1α、P-gp蛋白在胰腺癌组织中的表达,探讨它们的临床病理学意义及其相互之间的相关性.方法 应用免疫组织化学法检测74例胰腺癌组织,10例正常胰腺组织中HIF-1α、P-gp蛋白的表达.结果 HIF-1α、P-gp蛋白在胰腺癌组织中的阳性表达率分别为75.7%和86.5%,而正常胰腺组织中均呈阴性表达(P＜0.05).胰腺癌组织中HIF-1α和P-gp蛋白的表达呈显著正相关(r=0.304,P=0.009).胰腺癌组织中的HIF-1α蛋白表达与淋巴结转移、TNM分期呈正相关(P＜0.05);胰腺癌组织中的P-gp蛋白表达与淋巴结转移呈正相关,P-gp高表达与年龄大于60岁有关(P＜0.05).HIF-1α和P-gp蛋白高表达者的中位生存期明显短于低表达者(P＜0.05).结论 胰腺癌组织中HIF-1α、P-gp蛋白的表达上调,HIF-1α和P-gp呈正相关.胰腺癌组织中HIF-1α和P-gp蛋白的表达与淋巴结转移呈正相关.胰腺癌组织中HIF-1α和P-gp蛋白的高表达与预后差有关.

Abstract：

Objective To investigate the expression of HIF-1α and P-gp protein in pancreatic carcinoma and determine their clinicopathological significance and the correlation between the expression of HIF-1α, P-gp and the clinical prognosis. Method In samples from 74 cases of pancreatic carcinoma and 10 healthy individuals, the expression of HIF-1α and P-gp were detected by immunohistochemical method. Results The positive expression rate of HIF-1α and P-gp was 75.7% and 86.5%,respectively, which were remarkably higher than that in normal pancreatic tissue (P＜0.05). There was a positive correlation between the expression of HIF-1α and that of P-gp. The aberrant expression of HIF-1α and P-gp was associated with lymph node metastasis but not the location, size, clinical stages and nerve invasion of the tumor. Patients with high intensity of HIF-1α and P-gp expression showed a significantly lower median survival time than those with low intensity expression.Conclusions The expression of HIF-1α and P-gp is up-regulated in pancreatic carcinoma and there is a positive correlation between them. The expression of HIF-1α and P-gp might be related to the lymph node metastasis and poor prognosis.     

Wnt/β-catenin通路及靶基因c-myc在肝细胞癌组织中的表达

目的 观察Wnt通路激活在肝细胞癌起源中的作用.方法 应用免疫组织化学方法检测31例肝细胞癌患者肿瘤组织(实验组)与癌旁正常组织(对照组)中Wnt2、β-catenin及其靶基因c-myc的表达.结果 在肿瘤组织与癌旁正常组织中,Wnt2蛋白的表达差异无统计学意义(P＞0.05);β-catenin的胞质表达在癌旁组织中明显增高(P＜0.05),而其胞核表达则在肿瘤组织中增高(P＜0.05);c-myc在肿瘤组织中表达增高,两组差异有统计学意义(P＜0.01).结论 β-catenin及其靶基因c-myc在肝细胞癌组织中表达增高,提示Wnt通路激活的过程在肝细胞癌的起源中起到重要作用.

Abstract：

Objective To explore the effect of Wnt signaling pathway activation in the origination of hepatocellular carcinoma. Methods Immunohistochemistry was used to observe the expression of Wnt2,β-catenin and target gene c-myc in hepatocellular carcinoma (experimental group) and peficancerous tissue (control group) from 31 patients. Results The expression of Wnt2 had no significantly difference in both groups (P ＞0. 05). The expression of β-catenin in cytoplasm was significantly higher in peficancerous tissue. However,the expression of β-catenin in nucleus was significantly higher in hepatocellular carcinoma ( P ＜ 0. 05). The expression of c-myc was also significantly higher in hepatocellular carcinoma (P ＜ 0. 01).Conclusion β-catenin and it' s target gene c-myc overexpressed in hepatocellular carcinoma. The activation of Wnt signaling pathway played an important role in the origination of hepatocellular carcinoma.     

Identification and differentiation of PDX1 β-cell progenitors within the human pancreatic epithelium

AIM:To minimize the expansion of pancreatic mesenchymal cells in vitro and confirm thatβ-cell progenitors reside within the pancreatic epithelium.METHODS:Due to mesenchymal stem cell(MSC)expansion and overgrowth,progenitor cells within the pancreatic epithelium cannot be characterized in vitro,thoughβ-cell dedifferentiation and expansion of MSC intermediates via epithelial-mesenchymal transition(EMT)may generateβ-cell progenitors.Pancreatic epithelial cells from endocrine and non-endocrine tissue were expanded and differentiated in a novel pancreatic epithelial expansion medium supplemented with growth factors known to support epithelial cell growth(dexamethasone,epidermal growth factor,3,5,3’-triiodo-l-thyronine,bovine brain extract).Cells were also infected with a single and dual lentiviral reporter prior to cell differentiation.Enhanced green fluorescent protein was controlled by the rat Insulin 1 promoter and the monomeric red fluorescent protein was controlled by the mouse PDX1 promoter.In combination with lentiviral tracing,cells expanded and differentiated in the pancreatic medium were characterized by flow cytometry(BD fluorescence activated cell sorting),immunostaining and real-time polymerase chain reaction(PCR)(7900HT Fast Realtime PCR System).RESULTS:In the presence of 10%serum MSCs rapidly expand in vitro while the epithelial cell population declines.The percentage of vimentin+cells increased from 22%±5.83%to 80.43%±3.24%(14 d)and99.00%±0.0%(21 d),and the percentage of epithelial cells decreased from 74.71%±8.34%to 26.57%±9.75%(14 d)and 4.00%±1.53%(21 d),P0.01 for all time points.Our novel pancreatic epithelial expansion medium preserved the epithelial cell phenotype and minimized epithelial cell dedifferentiation and EMT.Cells expanded in our epithelial medium contained significantly less mesenchymal cells(vimentin+)compared to controls(44.87%±4.93%vs 95.67%±1.36%;P0.01).During cell differentiation lentiviral reporting demonstrated that,PDX1+and insulin+cells were localized within adherent epithelial cell aggregates compared to controls.Compared to starting islets differentiated cells had at least two fold higher gene expression of PDX1,insulin,PAX4 and RFX(P0.05).CONCLUSION:PDX1+cells were confined to adherent epithelial cell aggregates and not vimentin+cells(mesenchymal),suggesting that EMT is not a mechanism for generating pancreatic progenitor cells.     

肾细胞癌组织中DLK1蛋白表达及与临床病理学特征的相关性研究

目的 探讨不同组织类型的肾细胞癌组织中DLK1蛋白表达与肾癌临床病理特征及转移的关系.方法 采用免疫组织化学方法,检测94例肾原发透明细胞癌、76例乳头状肾细胞癌、45例嫌色细胞癌、71例透明细胞癌远处转移灶、24例透明细胞癌淋巴结转移灶及18例正常肾组织标本中DLK1蛋白表达情况,分析其与临床病理特征及转移的相关性.结果 正常肾组织近端及远端肾小管中DLK1蛋白表达均为阳性,透明细胞癌、乳头状肾细胞癌及嫌色细胞癌组织低表达率分别为33.0%(31/94)、27.6%(21/76)及33.3%(15/45),与正常肾组织相比,差异有统计学意义(P＜0.05),3种类型肾癌组织之间DLK1蛋白表达差异无统计学意义(P＞0.05).DLK1蛋白表达水平与透明细胞癌患者性别(男60例,女34例)、年龄(≥55岁50例,＜55岁44例)、病理分期(Ⅰ期41例,Ⅱ期9例,Ⅲ期21例,Ⅳ期23例)及淋巴结转移状态(无转移76例,有转移18例)等无明显相关性(P＞0.05).透明细胞癌原发灶、淋巴结转移灶、远处转移灶肿瘤组织中DLK1蛋白表达差异无统计学意义(P＞0.05).结论 不同类型的肾细胞癌组织中DLK1蛋白表达降低.DLK1蛋白低表达与透明细胞癌的临床病理特征及肿瘤转移无关.

Abstract：

Objective To identify the expression of DLK1 protein in different types of renal cell carcinomas and its correlations with pathological characteristics and metastasis. Methods Immunohistochemistry analysis was performed to evaluate the expression of DLK1 protein in 94 cases of primary clear cell renal cell carcinoma, 76 cases of papillary renal cell carcinoma, 45 cases of chromophobe renal cell carcinoma, 71 cases of distal metastatic and 24 cases of lymph node metastatic clear cell renal cell carcinoma, as well as 18 cases of normal renal tissue. The correlations of DLK1 protein expression with pathological characteristics were analyzed. Results DLK1 protein was expressed in proximal and distal renal tubular epithelial cells in all the normal renal cases. In contrast, DLK1 protein expression was lower in different types of renal cell carcinoma. The low or negative expression of DLK1 protein in clear cell renal cell carcinoma, papillary renal cell carcinoma and chromophobe renal cell carcinoma was 33.0% (31/94), 27.6% (21/76) and 33.3% (15/45), respectively. Compared to normal renal tissue, DLK1 protein expression was significantly down-regulated in renal cell carcinomas (P＞0.05), whereas there was no significant difference on DLK1 protein expressions among the different types (P＞0.05) of renal cell carcinomas. DLK1 protein expression was not correlated with sex (60 male and 34 female cases), age (≥55, 50 cases and 55, 44 cases), grade (41 cases in grade I, 9 cases in grade II, 21 cases in grade III and 23 cases in grade Ⅳ respectively) and lymph node metastasis (76 cases with and 18 cases without lymph node metastasis) in clear cell renal cell carcinoma (P＞0.05). There was also no significant difference among primary, lymph node and distal metastatic lesions of clear cell carcinoma (P＞0.05). Conclusions DLK1 protein expression is commonly down-regulated in different types of renal cell carcinomas. Down-regulation of DLK1 protein expression is not associated with pathological characteristics and metastasis in clear cell renal cell carcinoma.     

上皮钙黏素1基因启动子甲基化对结肠癌上皮钙黏素和β-连接素表达的影响

目的 探讨上皮钙黏素基因(CDH1)启动子甲基化与结肠癌上皮钙黏素(E-cadherin)及β-连接素(β-catenin)的表达及临床病理特征的关系.方法 采用甲基化特异性PCR技术检测68例结肠腺癌组织、癌旁组织及正常黏膜组织中CDH1基因启动子甲基化的状况.采用免疫组织化学法检测E-cadherin及β-catenin蛋白的表达.结果 癌旁组织及癌组织中CDH1启动子甲基化的阳性表达分别为32.4%(22/68)、57.4%(39/68),正常组织均为阴性表达(P<0.05).E-cadherin在正常组织、癌旁组织及腺癌组织中阳性表达率分别为92.6%、66.2%和44.1%.正常组织中β-catenin均表达于细胞膜上,无胞质和(或)胞核表达,而β-catenin在癌旁组织及癌组织中胞质和(或)胞核表达分别为29.4%和50.0%.CDH1基因启动子甲基化阳性率与E-cadherin表达则呈负相关(r=-0.312,P=0.01),与β-catenin胞质和(或)胞核表达呈正相关(r=0.309,P=0.018).CDH1基因启动子甲基化及E-cadherin、β-catenin的异常表达均与结肠癌分化程度及转移密切相关(P<0.05).结论 CDH1基因启动子甲基化可能是导致结肠癌E-cadherin与β-catenin异常表达及肿瘤侵袭性增强的重要原因.

Abstract：

Objective To investigate the relationship between methylation of the CDH1 gene promoter on the expression of E-cadherin and β-catenin, and to evaluate the correlation with clinicopathological characteristics of the colonic carcinoma. Methods Methylation specific PCR (MSP) was used to detect CDH1 gene promoter methylation in the cancer tissue, adjacent tissues and normal tissues in 68 patients. The expression of E-cadherin and β-catenin was determined by immunohistochemistry staining. Results The positive rate of CDH1 gene promoter methylation was 32.4% in adjacent tissues and 57.4% in cancer tissue, while no detectable methylation was found in all the normal tissues. The difference was statistically significant. The positive rate of E-cadherin was 92.6% in the normal tissues, 66.2% in the adjacent tissues and 44.1% in the cancer tissues. In all normal tissues, β-catenin was expressed only at the cellular membrane but not in the cytosol or nucleus, while the expression of β-catenin was present in the cytosol or nucleus in 29.4% of the adjacent tissues and 50.0% of the cancer tissues. The positive rate of CDH1 gene promoter methylation was negatively correlated with E-cadherin expression (r =-0.312,P =0.01) and positively correlated with β-catenin cytosolic/nucleus expression(r=0.309,P=0.018). The differentiation and metastasis of colonic carcinoma were associated with the aberrant expression of E-cadherin, β-catenin, and methylation of CDH1 promoter (P<0.05). Conclusion CDH1 gene promoter methylation may lead to aberrant expression of E-cadherin and β-catenin in colonic carcinoma, and may play an important role in promoting the invasion of tumor.     

肝卵圆细胞对肝纤维化大鼠细胞外信号调节蛋白和丝裂原激活蛋白激酶的影响

目的 观察肝卵圆细胞(hepatic oval cell,HOC)对肝纤维化(hepatofibrosis,HF)大鼠肝组织中细胞外信号调节蛋白(extracellular regulated protein kinases,ERK)和丝裂原激活蛋白激酶(mitogen activated protein kinase,P38MAPK)信号通路蛋白的影响.方法 SD大鼠以高脂低蛋白饲养、饮用10%乙醇,皮下注射40%CCl4,隔4 d注射一次,连续8周制备HF模型.HF大鼠采用胶原酶原位灌注法分离,percall纯化原代HOC.HOC悬液0.5 ml(1×109个细胞)经门静脉植入8周HF大鼠的肝脏内,分别于8、15、30d处死大鼠,HE和Masson染色观察HF病理组织学变化,Western blot 法检测肝组织ERK与P38MAPK信号通路蛋白的表达,同时检测ALB、FGF-3、c-kit、HNF-1α、PCNA表达.结果 病理组织学显示,HOC处理组大鼠HF被部分逆转,肝组织胶原纤维增生程度明显减轻,肝组织Ras、ERK、p-ERK、c-fos、c-jun、STAT3、ALB、FGF-3、PCNA蛋白表达显著下降(分别F=91.88,36.28,54.66,93.07,64.76,58.49,52.63,20.45,27.03,均P＜0.05),而HNF-α1和c-kit表达上调(分别F=18.63,25.99,均P＜0.05).结论 HOC抑制HF活化的ERK信号通路而改善肝纤维化程度,在HOC存在条件下p-P38并不激活c-fos、c-jun、STAT3、5表达,c-kit和HNF-1α表达增加,而肝组织结构损害程度明显减轻,肝纤维化程度明显改善.

Abstract：

Objective To observe the influence of hepatic oval cell (HOC) on the expression ERK and P38MAPK signaling pathway protein in liver tissue of murine experimental hepatofibrosis (HF).Method SD rats were fed with 10% ethanol and food with high-fat and low-protein, and were injected subcutaneously with carbontetrachloride once every four days for 8 weeks to establish hepatic fibrosis. HOGs were isolated from male HF rats by collagenase porfusion of the liver. HF rats at 8th week were transplanted with 0. 5 ml HOC suspension medium at a density of 1 × 109 cell /ml via portal vein, and the rats were sacrificed at 8th, 15th, 30th day respectively. Histopathologic changes of liver tissues were observed by HE and Masson. The expression of ERK and P38MAPK signaling pathway protein were determined by Western blotting. Result Hepatofibrosis was reversed and the degree of hyperplasia fibrilcollagen in hepatic fibrosis rats decreased significantly by HOC transplantion. HOC down-regulated the protein expression of Ras, ERK,p-ERK, c-fos, c-jun, STAT3, ALB, FGF-3, PCNA ( F = 91.88,36.28,54.66,93.07,64.76,58.49,52.63,20.45 ,27.03, all P ＜ 0.05 ), up-regulated the protein expression level of HNF-α1, PDGF-Rβ significantly in liver tissues(F = 18.63,25.99,P ＜0.05). Conclusions HOC improves the degree of hepatofibrosis through inhibiting hyperplasia of collagen fibril in liver tissue of hepatofibrosis rats. With the presence of HOC the expression of c-fos,c-jun,STAT3,5 was not activated by p-P38MAPK. The expression of c-kit and HNF-1α increased and that liver tissue injury alleviated, and hepatofibrosis was improved.     

成熟与未成熟心肌雌激素受体α和β的表达及其意义

目的研究大鼠成熟与未成熟心肌雌激素受体（ER）α和β的表达及其意义。方法采用半定量逆转录聚合酶链反应和免疫荧光组织化学方法，测定新生和成年大鼠心脏各部的ERα和ERβ mRNA及其蛋白质的表达。结果半定量RT-PCR分析显示ERα mRNA在未成熟心肌表达较低，但在成熟心肌ERα mRNA的表达水平显著提高。相反，未成熟心肌ERβ mRNA表达水平较高，而成熟心肌ERβ mRNA表达水平明显降低。免疫荧光组织化学图像分析证实这种差别也体现在雌激素受体α和β的蛋白质表达上。结论在成熟和未成熟心肌雌激素受本α和β并存，但存在明显的表达差异；ERα是雌激素调控成熟心肌的优势受体，在稳定成熟心肌的基因表达型发挥重要作用；而ERβ在未成熟心肌细胞的生长发育过程中起主导作用。